GEO database at the National Center for Biotechnology
Information and European Bioinformatics Institute (EBI) ArrayExpress
follow the MIAME format for data storage. Minimum Information About a
Microarray Experiment (MIAME) is a set of guidelines developed by the
Microarray Gene Expression Data (MGED). It been adopted by many journals
to support an independent evaluation of results.
Among the several R packages that can be used to
access its data, GEOquery is a simple and widely used one.
To start, we will need
if (!require("BiocManager", quietly = TRUE)) install.packages("BiocManager")
BiocManager::install("GEOquery", force = TRUE)library(GEOquery)
library(tidyverse)
library(pryr)The main user-level function in the GEOquery package is “getGEO”
args(getGEO)## function (GEO = NULL, filename = NULL, destdir = tempdir(), GSElimits = NULL,
## GSEMatrix = TRUE, AnnotGPL = FALSE, getGPL = TRUE, parseCharacteristics = TRUE)
## NULL
To download the data to a specific folder, we will need to create one. We could also provide an exisiting location.
# To store the data to specified folder, we need the the Dataset,
dir.create("GeoData")We will access the data submitted in the following article,
“Recurrent somatic mutations of PTPN1 in primary
mediastinal B cell lymphoma and Hodgkin lymphoma”. Nat Genet 2014
Apr;46(4):329-35
gds <- GEOquery::getGEO(GEO = "GDS5288", destdir = "GeoData")Meta() returns a list of object metadata
GEOquery::Meta(gds)[c("title", "description", "ref", "platform", "platform_technology_type",
"sample_organism", "sample_id", "sample_type")]## $title
## [1] "PTPN1 depletion effect on Hodgkin lymphoma derived cell line"
##
## $description
## [1] "Analysis of KM-H2 Hodgkin lymphoma derived cells depleted for PTPN1. PTPN1 encodes PTP1B, a non-receptor-type member of the superfamily of protein tyrosine phosphatases. Results provide insight into the role of PTPN1 mutations in lymphomagenesis."
## [2] "PTPN1 depletion"
## [3] "control"
##
## $ref
## [1] "Nucleic Acids Res. 2005 Jan 1;33 Database Issue:D562-6"
##
## $platform
## [1] "GPL570"
##
## $platform_technology_type
## [1] "in situ oligonucleotide"
##
## $sample_organism
## [1] "Homo sapiens"
##
## $sample_id
## [1] "GSM1308977,GSM1308978" "GSM1308975,GSM1308976"
##
## $sample_type
## [1] "RNA"
Columns() returns a data.frame of column descriptors. This includes information about each of the samples in the data.
GEOquery::Columns(gds)## sample protocol
## 1 GSM1308977 PTPN1 depletion
## 2 GSM1308978 PTPN1 depletion
## 3 GSM1308975 control
## 4 GSM1308976 control
## description
## 1 Value for GSM1308977: KM-H2 PTPN1 Knockdown, technical rep1; src: KM-H2 with PTPN1 siRNA knockdown
## 2 Value for GSM1308978: KM-H2 PTPN1 Knockdown, technical rep2; src: KM-H2 with PTPN1 siRNA knockdown
## 3 Value for GSM1308975: KM-H2 Non-silencing, technical rep1; src: KM-H2 non-silencing
## 4 Value for GSM1308976: KM-H2 Non-silencing, technical rep2; src: KM-H2 non-silencing
Table() returns a data.frame of gene expression data. The columns include gene IDs and expression values by Sample.
GEOquery::Table(gds)[1:5, ]## ID_REF IDENTIFIER GSM1308977 GSM1308978 GSM1308975 GSM1308976
## 1 1007_s_at MIR4640 8.82235 8.86388 8.58477 8.72635
## 2 1053_at RFC2 10.03960 9.81700 10.08130 9.80014
## 3 117_at HSPA6 5.98696 6.20082 6.18230 6.70773
## 4 121_at PAX8 7.74991 8.02246 7.94855 8.20815
## 5 1255_g_at GUCA1A 4.10924 3.83838 3.88709 4.00901
pryr::object_size(gds)## 7.86 MB
class(gds)## [1] "GDS"
## attr(,"package")
## [1] "GEOquery"
isS4(gds)## [1] TRUE
There are similar workflows for Sample, Series and Platform.
gsm <- getGEO("GSM1308977", destdir = "GeoData")Meta(gsm)## $channel_count
## [1] "1"
##
## $characteristics_ch1
## [1] "tissue: Hodgkin lymphoma" "cell line: KM-H2"
##
## $contact_address
## [1] "675 West 10th Avenue"
##
## $contact_city
## [1] "Vancouver"
##
## $contact_country
## [1] "Canada"
##
## $contact_department
## [1] "Pathology"
##
## $contact_email
## [1] "csteidl@bccancer.bc.ca"
##
## $contact_institute
## [1] "BC Cancer Agency"
##
## $contact_laboratory
## [1] "Steidl lab"
##
## $contact_name
## [1] "Christian,,Steidl"
##
## $contact_phone
## [1] "1-604-675-8046"
##
## $contact_state
## [1] "BC"
##
## $`contact_zip/postal_code`
## [1] "V5Z 1L3"
##
## $data_processing
## [1] "The R package aroma.affymetrix (v2.9.0) from Bioconductor was used to generate RMA expression and perform QC."
##
## $data_row_count
## [1] "54675"
##
## $extract_protocol_ch1
## [1] "Qiagen Allprep extraction columns"
##
## $geo_accession
## [1] "GSM1308977"
##
## $growth_protocol_ch1
## [1] "Cell lines: Standard conditions (www.dsmz.de)"
##
## $hyb_protocol
## [1] "Following fragmentation, cRNA were hybridized O/N at 45C on Affymetrix GeneChip HG 2.0 Plus Arrays. GeneChips were washed and stained in the Affymetrix Fluidics Station 450."
##
## $label_ch1
## [1] "biotin"
##
## $label_protocol_ch1
## [1] "Biotinylated cRNA were prepared according to the standard Affymetrix protocol (2-cycle amplification)"
##
## $last_update_date
## [1] "Jan 17 2014"
##
## $molecule_ch1
## [1] "total RNA"
##
## $organism_ch1
## [1] "Homo sapiens"
##
## $platform_id
## [1] "GPL570"
##
## $scan_protocol
## [1] "GeneChips were scanned using Affymetrix GeneChip Scanner"
##
## $series_id
## [1] "GSE54157"
##
## $source_name_ch1
## [1] "KM-H2 with PTPN1 siRNA knockdown"
##
## $status
## [1] "Public on Jan 17 2014"
##
## $submission_date
## [1] "Jan 16 2014"
##
## $supplementary_file
## [1] "ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM1308nnn/GSM1308977/suppl/GSM1308977_KMH2-PTPN1KD1.CEL.gz"
##
## $taxid_ch1
## [1] "9606"
##
## $title
## [1] "KM-H2 PTPN1 Knockdown, technical rep1"
##
## $type
## [1] "RNA"
Similar accessor functions work
Columns(gsm)## Column Description
## 1 ID_REF
## 2 VALUE log2 RMA normalized signal
Table(gsm)## ID_REF VALUE
## 1 AFFX-BioB-5_at 8.480296
## 2 AFFX-BioB-M_at 9.087702
## 3 AFFX-BioB-3_at 8.707758
## 4 AFFX-BioC-5_at 9.845029
## 5 AFFX-BioC-3_at 10.415414
## 6 AFFX-BioDn-5_at 11.051690
## 7 AFFX-BioDn-3_at 12.035703
## 8 AFFX-CreX-5_at 13.612359
## 9 AFFX-CreX-3_at 13.632270
## 10 AFFX-DapX-5_at 7.975419
## 11 AFFX-DapX-M_at 10.315585
## 12 AFFX-DapX-3_at 10.890056
## 13 AFFX-LysX-5_at 6.740159
## 14 AFFX-LysX-M_at 7.525384
## 15 AFFX-LysX-3_at 7.924963
## 16 AFFX-PheX-5_at 7.571823
## 17 AFFX-PheX-M_at 7.816444
## 18 AFFX-PheX-3_at 7.906589
## 19 AFFX-ThrX-5_at 6.523586
## 20 AFFX-ThrX-M_at 7.921625
## 21 AFFX-ThrX-3_at 9.298156
## 22 AFFX-TrpnX-5_at 3.860871
## 23 AFFX-TrpnX-M_at 3.432896
## 24 AFFX-TrpnX-3_at 3.375616
## 25 AFFX-r2-Ec-bioB-5_at 8.921402
## 26 AFFX-r2-Ec-bioB-M_at 9.382877
## 27 AFFX-r2-Ec-bioB-3_at 9.190695
## 28 AFFX-r2-Ec-bioC-5_at 10.524649
## 29 AFFX-r2-Ec-bioC-3_at 10.951632
## 30 AFFX-r2-Ec-bioD-5_at 12.445226
## 31 AFFX-r2-Ec-bioD-3_at 12.661747
## 32 AFFX-r2-P1-cre-5_at 14.112683
## 33 AFFX-r2-P1-cre-3_at 14.307971
## 34 AFFX-r2-Bs-dap-5_at 8.819005
## 35 AFFX-r2-Bs-dap-M_at 10.671276
## 36 AFFX-r2-Bs-dap-3_at 11.319084
## 37 AFFX-r2-Bs-lys-5_at 7.539878
## 38 AFFX-r2-Bs-lys-M_at 7.334497
## 39 AFFX-r2-Bs-lys-3_at 7.483875
## 40 AFFX-r2-Bs-phe-5_at 7.930399
## 41 AFFX-r2-Bs-phe-M_at 8.124227
## 42 AFFX-r2-Bs-phe-3_at 8.738725
## 43 AFFX-r2-Bs-thr-3_s_at 9.716796
## 44 AFFX-r2-Bs-thr-M_s_at 8.520607
## 45 AFFX-r2-Bs-thr-5_s_at 6.876051
## 46 AFFX-HUMISGF3A/M97935_5_at 6.641523
## 47 AFFX-HUMISGF3A/M97935_MA_at 7.896011
## 48 AFFX-HUMISGF3A/M97935_MB_at 8.464774
## 49 AFFX-HUMISGF3A/M97935_3_at 10.621345
## 50 AFFX-HUMRGE/M10098_5_at 13.464086
## 51 AFFX-HUMRGE/M10098_M_at 12.116453
## 52 AFFX-HUMRGE/M10098_3_at 11.691937
## 53 AFFX-HUMGAPDH/M33197_5_at 13.722883
## 54 AFFX-HUMGAPDH/M33197_M_at 13.912943
## 55 AFFX-HUMGAPDH/M33197_3_at 14.015626
## 56 AFFX-HSAC07/X00351_5_at 11.537511
## 57 AFFX-HSAC07/X00351_M_at 12.292272
## 58 AFFX-HSAC07/X00351_3_at 12.864160
## 59 AFFX-M27830_5_at 12.512481
## 60 AFFX-M27830_M_at 9.820761
## 61 AFFX-M27830_3_at 5.920335
## 62 AFFX-hum_alu_at 14.461802
## 63 1007_s_at 8.822346
## 64 1053_at 10.039576
## 65 117_at 5.986958
## 66 121_at 7.749908
## 67 1255_g_at 4.109242
## 68 1294_at 5.745199
## 69 1316_at 6.573133
## 70 1320_at 4.426695
## 71 1405_i_at 11.308156
## 72 1431_at 3.689785
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## 49920 1559988_at 4.143478
## 49921 1559989_at 5.599989
## 49922 1559990_at 3.142747
## 49923 1559991_s_at 4.583763
## 49924 1559992_a_at 2.386535
## 49925 1559993_at 7.284075
## 49926 1559995_at 5.448935
## 49927 1559996_s_at 5.391509
## 49928 1559997_x_at 5.300716
## 49929 1560001_at 4.840550
## 49930 1560002_at 4.735466
## 49931 1560006_a_at 5.272466
## 49932 1560007_at 4.039347
## 49933 1560010_a_at 4.217594
## 49934 1560011_at 4.613690
## 49935 1560012_at 3.148340
## 49936 1560013_at 5.157034
## 49937 1560014_s_at 5.256121
## 49938 1560017_at 5.966949
## 49939 1560018_at 2.869174
## 49940 1560019_at 3.404327
## 49941 1560020_at 4.195645
## 49942 1560021_at 4.345997
## 49943 1560023_x_at 4.911993
## 49944 1560024_at 4.369460
## 49945 1560025_at 4.840281
## 49946 1560026_at 4.451138
## 49947 1560028_at 3.591151
## 49948 1560029_a_at 3.481976
## 49949 1560030_at 3.894180
## 49950 1560031_at 3.860081
## 49951 1560033_at 4.030683
## 49952 1560034_a_at 3.343461
## 49953 1560035_at 5.199617
## 49954 1560038_at 3.214297
## 49955 1560042_at 4.226645
## 49956 1560043_at 4.126209
## 49957 1560047_s_at 4.874019
## 49958 1560048_at 3.231632
## 49959 1560049_at 4.679500
## 49960 1560051_at 3.058721
## 49961 1560058_at 4.874788
## 49962 1560059_at 5.838865
## 49963 1560060_s_at 9.751424
## 49964 1560062_at 4.981271
## 49965 1560063_a_at 3.182951
## 49966 1560065_at 4.251447
## 49967 1560066_at 5.158509
## 49968 1560068_a_at 4.347552
## 49969 1560069_at 4.230301
## 49970 1560070_at 3.961065
## 49971 1560071_a_at 5.658801
## 49972 1560074_at 4.915658
## 49973 1560075_at 3.222060
## 49974 1560078_at 3.854081
## 49975 1560080_at 3.110333
## 49976 1560081_at 5.299379
## 49977 1560082_at 4.676392
## 49978 1560086_at 2.942020
## 49979 1560087_a_at 3.834042
## 49980 1560089_at 6.600788
## 49981 1560094_at 4.548987
## 49982 1560095_s_at 4.667635
## 49983 1560097_at 5.053717
## 49984 1560098_at 2.948821
## 49985 1560099_at 6.904948
## 49986 1560100_at 5.560983
## 49987 1560101_at 4.914911
## 49988 1560102_at 4.518731
## 49989 1560104_at 3.193353
## 49990 1560105_at 3.620768
## 49991 1560107_at 2.619142
## 49992 1560108_at 3.911528
## 49993 1560109_s_at 4.385759
## 49994 1560111_at 2.472636
## 49995 1560112_at 3.170540
## 49996 1560115_a_at 5.695858
## 49997 1560116_a_at 10.109542
## 49998 1560117_at 4.449395
## 49999 1560118_at 3.519767
## [ reached 'max' / getOption("max.print") -- omitted 4676 rows ]
pryr::object_size(gsm)## 4.39 MB
class(gsm)## [1] "GSM"
## attr(,"package")
## [1] "GEOquery"
isS4(gsm)## [1] TRUE
Here is obtaining GPL metadata.
gpl <- getGEO("GPL570", destdir = "GeoData")pryr::object_size(gpl)## 74.01 MB
class(gpl)## [1] "GPL"
## attr(,"package")
## [1] "GEOquery"
Meta(gpl) %>%
str(vec.len = 1, nchar.max = 65)## List of 27
## $ contact_address : chr " "
## $ contact_city : chr "Santa Clara"
## $ contact_country : chr "USA"
## $ contact_email : chr "geo@ncbi.nlm.nih.gov, support@affymetrix.com"
## $ contact_institute : chr "Affymetrix, Inc."
## $ contact_name : chr " ,,Affymetrix, Inc."
## $ contact_phone : chr "888-362-2447"
## $ contact_state : chr "CA"
## $ contact_web_link : chr "http://www.affymetrix.com/index.affx"
## $ contact_zip/postal_code: chr "95051"
## $ data_row_count : chr "54675"
## $ description : chr [1:5] "Affymetrix submissions are typically submitted t"| __truncated__ ...
## $ distribution : chr "commercial"
## $ geo_accession : chr "GPL570"
## $ last_update_date : chr "Dec 14 2020"
## $ manufacture_protocol : chr [1:6] "see manufacturer's web site" ...
## $ manufacturer : chr "Affymetrix"
## $ organism : chr "Homo sapiens"
## $ relation : chr [1:61] "Alternative to: GPL4454 (Alternative CDF)" ...
## $ sample_id : chr [1:166727] "GSM18422" ...
## $ series_id : chr [1:5550] "GSE1145" ...
## $ status : chr "Public on Nov 07 2003"
## $ submission_date : chr "Nov 07 2003"
## $ taxid : chr "9606"
## $ technology : chr "in situ oligonucleotide"
## $ title : chr "[HG-U133_Plus_2] Affymetrix Human Genome U133 Plus 2.0 Array"
## $ web_link : chr [1:2] "http://www.affymetrix.com/support/technical/bypr"| __truncated__ ...
Columns(gpl) %>%
as_tibble()## # A tibble: 16 × 2
## Column Description
## <chr> <chr>
## 1 ID "Affymetrix Probe Set ID LINK_PRE:\"https:/…
## 2 GB_ACC "GenBank Accession Number LINK_PRE:\"http:/…
## 3 SPOT_ID "identifies controls"
## 4 Species Scientific Name "The genus and species of the organism repr…
## 5 Annotation Date "The date that the annotations for this pro…
## 6 Sequence Type ""
## 7 Sequence Source "The database from which the sequence used …
## 8 Target Description ""
## 9 Representative Public ID "The accession number of a representative s…
## 10 Gene Title "Title of Gene represented by the probe set…
## 11 Gene Symbol "A gene symbol, when one is available (from…
## 12 ENTREZ_GENE_ID "Entrez Gene Database UID LINK_PRE:\"http:/…
## 13 RefSeq Transcript ID "References to multiple sequences in RefSeq…
## 14 Gene Ontology Biological Process "Gene Ontology Consortium Biological Proces…
## 15 Gene Ontology Cellular Component "Gene Ontology Consortium Cellular Componen…
## 16 Gene Ontology Molecular Function "Gene Ontology Consortium Molecular Functio…
Table(gpl) %>%
dim()## [1] 54675 16
Table(gpl) %>%
as_tibble() %>%
dplyr::select(ID, GB_ACC, `Sequence Source`, `Target Description`)## # A tibble: 54,675 × 4
## ID GB_ACC `Sequence Source` `Target Description`
## <chr> <chr> <chr> <chr>
## 1 1007_s_at U48705 Affymetrix Proprietary Database U48705 /FEATURE=mRNA /DEFIN…
## 2 1053_at M87338 GenBank M87338 /FEATURE= /DEFINITIO…
## 3 117_at X51757 Affymetrix Proprietary Database X51757 /FEATURE=cds /DEFINI…
## 4 121_at X69699 GenBank X69699 /FEATURE= /DEFINITIO…
## 5 1255_g_at L36861 Affymetrix Proprietary Database L36861 /FEATURE=expanded_cd…
## 6 1294_at L13852 GenBank L13852 /FEATURE= /DEFINITIO…
## 7 1316_at X55005 Affymetrix Proprietary Database X55005 /FEATURE=mRNA /DEFIN…
## 8 1320_at X79510 Affymetrix Proprietary Database X79510 /FEATURE=cds /DEFINI…
## 9 1405_i_at M21121 GenBank M21121 /FEATURE= /DEFINITIO…
## 10 1431_at J02843 Affymetrix Proprietary Database J02843 /FEATURE=cds /DEFINI…
## # … with 54,665 more rows
gse <- getGEO(GEO = "GSE54157", destdir = "GeoData", GSEMatrix = FALSE) # Note the GSEMatrix argument
gse_exprs <- getGEO(GEO = "GSE54157", destdir = "GeoData", GSEMatrix = TRUE) # Note the GSEMatrix argumentclass(gse)## [1] "GSE"
## attr(,"package")
## [1] "GEOquery"
Meta(gse)[c("title", "type", "platform_id", "summary", "supplementary_file")]## $title
## [1] "Expression changes induced by PTPN1 knockdown in KM-H2 cells"
##
## $type
## [1] "Expression profiling by array"
##
## $platform_id
## [1] "GPL570"
##
## $summary
## [1] "Gene expression data for shRNA PTPN1 knockdown vs. Non-silencing in the classical Hodgkin lymphoma-derived cell line KM-H2"
##
## $supplementary_file
## [1] "ftp://ftp.ncbi.nlm.nih.gov/geo/series/GSE54nnn/GSE54157/suppl/GSE54157_RAW.tar"
We can extract out the GSM and GPL in the series.
GEOquery::GSMList(gse) %>%
names()## [1] "GSM1308975" "GSM1308976" "GSM1308977" "GSM1308978"
GEOquery::GPLList(gse) %>%
names()## [1] "GPL570"
gse_exprs <- getGEO(GEO = "GSE54157", destdir = "geoData", GSEMatrix = TRUE) # Note the GSEMatrix argumentclass(gse_exprs)## [1] "list"
length(gse_exprs)## [1] 1
pryr::object_size(gse_exprs)## 63.81 MB
lapply(gse_exprs, class)## $GSE54157_series_matrix.txt.gz
## [1] "ExpressionSet"
## attr(,"package")
## [1] "Biobase"
Phenotype data can be extracted from the ExpressionSet
gse_GPL570 <- gse_exprs[[1]]
Biobase::pData(gse_GPL570) %>%
as_tibble()In this case, the GEO series does not have pre-processed expression data. Instead, raw data can be accessed from the supplemental file links. Here we get two files. The full path to the download is saved as a rowname.
supp_info <- GEOquery::getGEOSuppFiles(GEO = "GSE147507")
rownames(supp_info)## [1] "/__w/RU_ReproducibleR_abridged/RU_ReproducibleR_abridged/extdata/GSE147507/GSE147507_RawReadCounts_Ferret.tsv.gz"
## [2] "/__w/RU_ReproducibleR_abridged/RU_ReproducibleR_abridged/extdata/GSE147507/GSE147507_RawReadCounts_Human.tsv.gz"
basename(rownames(supp_info)[1])## [1] "GSE147507_RawReadCounts_Ferret.tsv.gz"
basename(rownames(supp_info)[2])## [1] "GSE147507_RawReadCounts_Human.tsv.gz"
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